采用溶液核磁共振波谱和等温滴定量热法相结合的混合策略来表征蛋白-纳米盘相互作用。
A hybrid strategy combining solution NMR spectroscopy and isothermal titration calorimetry to characterize protein-nanodisc interaction.
机构信息
Institute for Protein Research, Osaka University, Yamadaoka, Suita, Osaka, 565-0871, Japan.
Institute for Protein Research, Osaka University, Yamadaoka, Suita, Osaka, 565-0871, Japan; Research Center for Bioconvergence Analysis, Korea Basic Science Institute, Chungcheongbuk-do, 28119, South Korea; Bio-Analytical Science, University of Science and Technology, Daejeon, 34113, South Korea; Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, 34134, South Korea.
出版信息
Anal Biochem. 2022 Feb 15;639:114521. doi: 10.1016/j.ab.2021.114521. Epub 2021 Dec 11.
NMR is a powerful tool for characterizing intermolecular interactions at atomic resolution. However, the nature of the complex interactions of membrane-binding proteins makes it difficult to elucidate the interaction mechanisms. Here, we demonstrated that structural and thermodynamic analyses using solution NMR spectroscopy and isothermal titration calorimetry (ITC) can clearly detect a specific interaction between the pleckstrin homology (PH) domain of ceramide transport protein (CERT) and phosphatidylinositol 4-monophosphate (PI4P) embedded in the lipid nanodisc, and distinguish the specific interaction from nonspecific interactions with the bulk surface of the lipid nanodisc. This NMR-ITC hybrid strategy provides detailed characterization of protein-lipid membrane interactions.
NMR 是一种强大的工具，可在原子分辨率下对分子间相互作用进行表征。然而，膜结合蛋白的复杂相互作用的性质使得阐明相互作用机制变得困难。在这里，我们证明了使用溶液 NMR 光谱和等温滴定量热法（ITC）进行的结构和热力学分析可以清楚地检测出神经酰胺转运蛋白（CERT）的pleckstrin 同源（PH）域与嵌入脂质纳米盘的磷脂酰肌醇 4-单磷酸（PI4P）之间的特异性相互作用，并将特异性相互作用与脂质纳米盘的体相非特异性相互作用区分开来。这种 NMR-ITC 混合策略提供了对蛋白质-脂质膜相互作用的详细表征。
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